Isolation and identification of an isoflavone reducing bacterium from feces from a pregnant horse.
Abstract: The aim of this research was to isolate bacteria capable of biotransforming daidzein from fresh feces from pregnant horses. A Hungate anaerobic roller tube was used for anaerobic culture. Single colonies were picked at random and incubated with daidzein. High performance liquid chromatography was used to detect whether the isolated bacteria were able to biotransform the substrate. A strain capable of reducing daidzein was selected and characterized using sequence analysis of 16S rDNA, and a phylogenetic tree was constructed. The morphological physiological and biochemical characteristics of the strain were investigated. A facultative anaerobic, Gram-positive bacterium capable of converting daidzein to dihydrodaidzein was isolated and named HXBM408 (MF992210). A BLAST search of HXBM408's 16S rDNA sequence against the GenBank database suggested that the strain has 99% similarity with Pediococcus acidilactici strain DSM (NR042057). The morphological, physiological, and biochemical characteristics of HXBM408 are very similar to those of Pediococcus. Based on these characteristics, the strain was identified as Pediococcus acidilactici. The bacterial strain HXBM408 isolated from the feces of pregnant horses was able to reduce the isoflavone daidzein to dihydrodaidzein.
Publication Date: 2019-11-18 PubMed ID: 31738752PubMed Central: PMC6860936DOI: 10.1371/journal.pone.0223503Google Scholar: Lookup
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- Journal Article
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Summary
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This research aimed to isolate and identify a specific bacterium able to biotransform a compound known as daidzein from the feces of pregnant horses. The identified bacterium, named HXBM408, was found to have a 99% genetic similarity with Pediococcus acidilactici and was capable of converting daidzein into a compound called dihydrodaidzein.
Methods and Technical Details
- The researchers used fresh feces from pregnant horses to gather their bacterial samples.
- An anaerobic culture technique, namely a Hungate anaerobic roller tube, was employed to allow the growth of bacteria that thrive in environments without oxygen.
- Isolated bacteria colonies were then cultured in the presence of daidzein, a compound they hoped the bacteria could convert or biotransform.
- They utilized high performance liquid chromatography, a technique used to distinguish, identify, and quantify each part of a mixture, to check whether the isolated bacteria successfully biotransformed daidzein.
Findings and Characterization
- Following the screening process, a strain that demonstrated the ability to reduce daidzein was selected and named HXBM408 (MF992210).
- The strain’s identity and characteristics were examined using sequence analysis of 16S rDNA, a highly conserved (i.e., minimal variance) region of DNA which is often used to identify and categorize bacteria.
- A phylogenetic tree, a graph demonstrating the evolutionary relationships between species, was generated based on the rDNA analysis results.
- Further analysis included studying the physiological, morphological, and biochemical characteristics of HXBM408.
- Results from this comprehensive characterization suggested that the bacterium was a Gram-positive, facultatively anaerobic strain, which means it can grow in either the presence or absence of oxygen.
Identification and Significance
- The researchers utilized a BLAST search against the GenBank database, a library of all publicly available genetic sequences, to compare HXBM408’s 16S rDNA sequence.
- The search indicated that HXBM408 shares 99% similarity with the strain Pediococcus acidilactici DSM (NR042057).
- Considering this high similarity, and that the physiological, morphological, and biochemical characteristics of HXBM408 corresponded to those typically seen in Pediococcus, the researchers identified HXBM408 as a strain of Pediococcus acidilactici.
- The successful identification of a bacterium capable of converting daidzein to dihydrodaidzein could have significant implications for understanding the role of gut flora in animal and human health, particularly relating to isoflavone absorption and metabolism.
Cite This Article
APA
Jinglong X, Xiaobin L, Fang Z, Chenchen W, Kailun Y.
(2019).
Isolation and identification of an isoflavone reducing bacterium from feces from a pregnant horse.
PLoS One, 14(11), e0223503.
https://doi.org/10.1371/journal.pone.0223503 Publication
Researcher Affiliations
- Xinjiang Laboratory of Meat-and Milk-Production Herbivore Nutrition, Xinjiang Agricultural University, Urumqi, Xinjiang, China.
- Xinjiang Laboratory of Meat-and Milk-Production Herbivore Nutrition, Xinjiang Agricultural University, Urumqi, Xinjiang, China.
- Xinjiang Laboratory of Meat-and Milk-Production Herbivore Nutrition, Xinjiang Agricultural University, Urumqi, Xinjiang, China.
- Xinjiang Laboratory of Meat-and Milk-Production Herbivore Nutrition, Xinjiang Agricultural University, Urumqi, Xinjiang, China.
- Xinjiang Laboratory of Meat-and Milk-Production Herbivore Nutrition, Xinjiang Agricultural University, Urumqi, Xinjiang, China.
MeSH Terms
- Animals
- Bacteria / genetics
- Bacteria / isolation & purification
- Bacteria / metabolism
- Biotransformation
- DNA, Bacterial / genetics
- DNA, Ribosomal / genetics
- Feces / microbiology
- Female
- Horses / metabolism
- Horses / microbiology
- Isoflavones / metabolism
- Pediococcus acidilactici / genetics
- Pediococcus acidilactici / isolation & purification
- Pediococcus acidilactici / metabolism
- Phylogeny
- Phytoestrogens / metabolism
- Pregnancy
- RNA, Ribosomal, 16S / genetics
- Soybeans / chemistry
Conflict of Interest Statement
The authors have declared that no competing interests exist.
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Citations
This article has been cited 3 times.- Okyere SK, Wen J, Cui Y, Xie L, Gao P, Zhang M, Wang J, Wang S, Ran Y, Ren Z, Hu Y. Bacillus toyonensis SAU-19 and SAU-20 Isolated From Ageratina adenophora Alleviates the Intestinal Structure and Integrity Damage Associated With Gut Dysbiosis in Mice Fed High Fat Diet. Front Microbiol 2022;13:820236.
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- Gong Y, Lv J, Pang X, Zhang S, Zhang G, Liu L, Wang Y, Li C. Advances in the Metabolic Mechanism and Functional Characteristics of Equol. Foods 2023 Jun 10;12(12).
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